Based of the study on the impact of protoplasts under different conditions of enzymolysis solution, culture time, enzymolysis time and enzymolysis temperature in sequence using orthogonal test. The best optimal condition of producing protoplast was :0.5% helicase add 0.5% lysozyme and 1.0% cellulase, enzymolysis time was 3 hours , enzymolysis temperature was 32℃,under these condition the production of protoplast can be 4.25?10~(6 )per mL. Ca~(2+) and PEG can increase the regeneration ratio of protoplast.After induced mutation by complex factor under DES,UV and LiCl,the mutagenesis strain which own increasing enzyme activity and transmissibility steaty was obtained.

Fusion PCR, which employs chimeric primers to generate PCR products with complementary ends in its amplifications, is a rapid and flexible method in joining different DNA fragments. This method can assemble DNA fragments without the treatment of restriction endonucleases and T4 DNA ligase. It offered a shortcut for the construction of homologous recombinant fragments. Through assembling three recombinant fragments successfully, fusion PCR procedure was improved and manipulation essentials of fusion PCR were described in detail. The results indicated that the improved procedure can assemble three or four fragments simultaneously, and the length of each fusion product is above 4.5 kb. The result recombinants were proposed to be use in further experiments, which structures had been confirmed by sequencing.

Based on a ?-tubulin gene EST sequence from Trichoderma harzianum mycelium cDNA library,a 1.74kb ?-tubulin gene coding region,together with a 1.5kb 5'-flanking region and a 1.0kb 3'-flanking region was cloned from T.harzianum by IPCR methods.The gene encodes a 446 amino acid polypeptide,which shows a high degree of homology with other fungal ?-tubulins.A three-dimensional model of the T.harzianum ?-tubulin was built based on the crystal structure of bovine tubulin using homology modeling method.Tertiary protein structure of the ?-tubulin should provide a basis for understanding a significant body of research on tubulin's properties and interactions with antimicrotubule agents.

Objective: To find the reason of risk on medical equipment. Methods: Based on RCA, to adopt the event classification to evaluate the X-ray equipment, high-frequency electro tome, electro-cardiac monitor, infusion pump, ventilator, defibrillators and infant incubator, discuss the root cause of medical risk. Results:To deduce the medical equipment risk by PM and quality control. Conclusion:With the analysis of quality control, the application of RCA is gradually explained in management of medical equipment.

BACKGROUND:It is confirmed that the onset of osteoarthritis is closely related to inflammatory cytokines. OBJECTIVE: To investigate the levels of interleukin-6 and matrix metaloproteinase-3 in the synovial fluid of patients with osteoarthritis and their correlation with the severity of osteoarthritis. METHODS: Synovial fluid specimens of 78 patients were harvested to detect the levels of interleukin-6 and matrix metaloproteinase-3 using ELISA assay. The severity of osteoarthritis was estimated by the Hospital for Special Surgery (HSS) score. Pearson correlation analysis was adopted to explore the pairwise correlations between interleukin-6, matrix metaloproteinase-3 and HSS score. RESULTS AND CONCLUSION:The levels of interleukin-6 and matrix metaloproteinase-3 in the synovial fluid were (13.1±5.7) and (989.3±429.5) μg/L, respectively; the HSS score was (56.4±12.0) points. Pearson correlation analysis showed that the expression levels of interleukin-6 and matrix metaloproteinase-3 had a significant positive correlation (r=0.70,P< 0.001); the levels of interleukin-6 and matrix metaloproteinase-3 had a negative correlation with the HSS scores (r=-0.70,r=-0.75,P < 0.001). Experimental findings show that there is a positive correlation between interleukin-6 and matrix metaloproteinase-3 in the synovial fluids, and these two cytokines also have a positive correlation with the severity of osteoarthritis, indicating that to measure interleukin-6 and matrix metaloproteinase-3 levels in the synovial fluids of osteoarthritis patients is important for early diagnosis and treatment of osteoarthritis.

Ras is a kind of protein closely related to neoplasms,the persistent activation of which would induce tumorigenesis and progression.RASAL1 is a Ca~(2+) regulated ras GTPase-activating-like protein, the loss or decreased expression of RASAL1 induces ras activation, then results in tumongenesis and progression.In this review we sum up the mechanism of ras activation, the relationship between neoplasms and RASAL1 and the mechanism of RASAL1 expression decreasing.

OBJECTIVE To investigate the infection and drug-resistance of Escherichia coli.METHODS Double disk diffusion test and improved three-dimensional test were adopted to test ESBLs and AmpC of 135 E.coli strains obtained from bile cultivation from operations of biliary inpatients in our hospital from 2003 to 2005,and King-Bauer′s was used to test the sensitivity of 14 kinds of common antibiotics.RESULTS The detection rate of ESBLs producing bacteria was 30.4%(41 strains),and that of AmpC producing bacteria was 5.2%(7 strains);drug-sensitivity test indicated,that E.coli was sensitive to meropenem,imipenem,cefoperazone / subactam,ciprofloxacin and amikacin.CONCLUSIONS E.coli is a main bacterium in biliary infection,and the strains producing ESBLs and /or AmpC are resistant to the cephalosporins of third and fourth generation;prematurely and excessively using broad-spectrum antibiotics before operation is the primary cause to increase in enzyme-producing strains.

In our university,the training for geriatric postgraduates of clinical medicine is now in the initial stage.Therefore,it is important to explore the training model to improve the quality of postgraduate.The program focused on cultivating geriatric postgraduates' ability of diagnosis and treatment,helping them have the concept of integral service and enhancing their teaching and thinking ability of clinical sciences to provide high-level comprehensive geriatric doctors for the society.

Objective To investigate and analyze the effect of psychological intervention combined with paclitaxel and carboplatin on the clinical efficacy and quality of life of locally advanced cervical cancer. Methods 100 cases of locally advanced cervical cancer treated in our hospital from February 2015 to November 2016 were selected and randomly divided into the control group and the experimental group, with 50 patients in each group. The control group were treated with neoadjuvant chemotherapy with cisplatin combined with fluorouracil, experimental group were treated with paclitaxel and carboplatin chemotherapy, and give attention to patients with psychological intervention, mental health, strengthen communication and exchanges, the elimination of patients with anxiety. The treatment effect and quality of life index of 2 groups were compared and analyzed. Results After treatment, the total score of quality of life in the experimental group was (75.2 ± 11.2) points, significantly higher than that of the control group (64.2 ± 9.3), with statistical difference (P<0.05). Among the patients in the experimental group, 5 patients progressed, and 9 patients were stable. The number of effective treatment was 36. In the control group, 10 patients progressed, 13 patients were stable, and the number of effective treatment was 27. Conclusion The clinical effect of psychological intervention combined with paclitaxel and carboplatin for local advanced cervical cancer is better, higher efficiency, can significantly improve the quality of life of patients, eliminate the negative emotion of patients, which has clinical significance.

Electrical status epilepticus in sleep (ESES) indicates a special electroencephalographic pattern showing sleep-induced continuous paroxysmal discharges of spike-wave complexes.ESES can be seen in a series of epileptic syndromes in children characterized by seizures,ESES and cognitive impairment,including epilepsy with continuous spikes and waves during slow sleep,Landau Kleffner syndrome,typical and atypical benign childhood epilepsy with centro-temporal spikes.Though the mechanism generating ESES remains elusive,great advances in several aspects,such as genetic studies (GRIN2A,copy number variations,ect.),high-frequency oscillations and brain networks have been achieved.Treatment for ESES related disorders should focus on both epileptic seizures and ESES,and the introduction of appropriate antiepileptic drugs or other strategies like hormone therapy should be considered to furthest eliminate epileptic seizures and protect cognitive function.